Chemiluminescent Immunoassay Research Articles

Development of a thrombin-antithrombin complex detection kit and study in venous thromboembolism complicated by cervical cancer

ObjectiveOur study successfully developed an assay kit for thrombin-antithrombin complex (TAT) and demonstrated the predictive value of plasma TAT concentration in the development of venous thromboembolism (VTE) in patients with cervical cancer.MethodA retrospective analysis was conducted on 177 patients with cervical cancer who received treatment at the Affiliated Hospital of Jiangnan University in Wuxi City from July 1, 2023 to October 1, 2023. This study provides a comprehensive analysis of cervical cancer patients and their VTE risk factors. The patients were divided into two groups: 27 cases with VTE (Thrombosis group) and 150 cases without VTE (Non-thrombotic group). Additionally, the patients were classified into four stages based on tumor stage: 42 cases of stage I, 45 cases of stage II, 62 cases of stage III, and 28 cases of stage IV. The control group consisted of 80 healthy patients undergoing medical check-ups. Thrombin-antithrombin complex (TAT), fibrinolytic enzyme-α2-fibrinolytic inhibitor complex (PIC), thrombomodulin (TM), and tissue-type plasminogen activator inhibitor 1 complex (t-PAIC) were detected using quantitative chemiluminescence immunoassay. The study assessed the variations in thrombotic marker levels among cervical cancer patients of different stages through a receiver operating characteristic (ROC) curve.ResultThe TAT reagent demonstrated a detection limit of 0.048 ng/mL, with a linear R value of 0.9997, indicating high accuracy and precision. The reagent's accelerated stability was also excellent, with deviations of less than 10%. Furthermore, the correlation coefficient of this method with Hyson Mecon was R2 = 0.9683. Notably, in patients with cervical cancer, TAT and PIC levels were found to be elevated compared to those of the healthy population. Cervical cancer patients who developed thrombosis had significantly elevated levels of TAT and fibrinogen degradation products (FDP) compared to those who did not. Furthermore, patients with stage III-IV cervical cancer exhibited higher levels of the six markers than those with stage I-II during staging. Notably, the combination of four or six markers significantly improved the sensitivity and specificity of the diagnosis, as demonstrated by the ROC curves.ConclusionOur developed TAT test kit has excellent performance and low cost, making it a clinically valuable tool for widespread use. Elevated TAT levels have significant predictive value for thrombosis occurrence in cervical cancer patients. The combination of t-PAIC, TM, TAT, PIC, D-dimer(D-D), and FDP markers is superior to using a single marker for diagnosing VTE in patients with malignant tumors. Screening cervical cancer patients for the six markers is essential to aid in active prophylaxis, determine optimal treatment timing, and implement nursing interventions to improve prognosis, reduce venous thrombosis incidence and mortality, and prolong survival time.

Association of Serum 25-Hydroxyvitamin D Level with the Grades of Hepatic Encephalopathy in Cirrhotic Patients

Background: Liver cirrhosis is the final phase of the hepatic fibrosis process, defined as a diffuse disruption of the normal architecture of the liver with fibrosis and nodule formation and it is one of the major cause of morbidity and mortality throughout the world. Hepatic encephalopathy (HE) is one of the complications of cirrhosis that identified in up to 55% of patients. The etiology of HE is complex and multifactorial. Vitamin D deficiency is more common in cirrhosis and associated with neuropsychiatric abnormalities and systemic inflammation as well as infection thereby giving rise to the development of HE. Objectives: This study aimed to evaluate the serum 25- Hydroxyvitamin D level in liver cirrhosis patient with or without hepatic encephalopathy and its relation with grades of hepatic encephalopathy. Materials and Methods: An observational cross-sectional study was carried out in the Department of Hepatology, Bangabandhu Sheikh Mujib Medical University, Dhaka from December 2019 to August 2020. Patients who met the selection criteria were counselled and finally 54 were included in the study among them 27 had hepatic encephalopathy where 27 had not. Serum 25-Hydroxyvitamin D level was estimated by chemiluminescence immunoassay (CLIA) in our Biochemistry department. All data were presented as mean ± SD & analyzed by SPSS (version 20). Qualitative data were analyzed by Chi-square test & quantitative data were analyzed by student’s t-test. Statistically significance of serum 25- Hydroxyvitamin D level among the grades of hepatic encephalopathy will be detected by ANOVA test. A statistically significant result was considered when p value less than 0.05. Result: Among 54 cirrhotic patients, age ranged from 20 to 72 years and the mean age was 50.2 ± 13.1 years. The Child-Pugh score and MELD score were negatively correlate with mean 25-Hydroxyvitamin D level. The mean level of 25-Hydroxyvitamin D ...

Detection of TNF-α using the established ab-MPs-CLIA

Tumor necrosis factor alpha (TNF-α) is a key cytokine in inflammation and immune responses, making its rapid and accurate detection essential for disease diagnosis and management. In this study, we developed a highly sensitive chemiluminescence immunoassay (CLIA) using antibody-coated magnetic particles (Ab-MPs-CLIA) for TNF-α detection. From nine candidate antibodies, we identified an optimal pair through epitope competition and affinity assessments, significantly improving assay performance. The Ab-MPs-CLIA achieved a detection limit of 0.25 pg/mL, 6.8 times more sensitive than Siemens commercial kits, with a broad linear range of 9.2–1077 pg/mL. The method demonstrated excellent stability, both under accelerated conditions at 37 °C for 7 days and long-term storage at 4 °C for 12 months. It showed no cross-reactivity with common interfering substances in human serum, ensuring high specificity. Notably, the entire process, from sample preparation to result, takes just 25 min, compared to 3–4 h for both ELISA and RIA, and CLIA typically offers 10–100 times higher sensitivity than these methods. These advantages make the Ab-MPs-CLIA an ideal option for clinical laboratories, providing superior sensitivity, specificity, broader dynamic range, and greater operational efficiency than existing TNF-α detection technologies.

Gold nanoclusters-incorporated nanogels as hybrid nanozymes for sensitive chemiluminescence immunoassays

Nanomaterials with excellent biocompatibility hold significant potential in chemiluminescence (CL) immunoassays. However, achieving green synthesis and maintaining the structural stability of such nanomaterials remain challenging. In this study, gold nanoclusters (Au NCs) were synthesized via the reduction of bovine serum albumin (BSA). These Au NCs were electrostatically combined with horseradish peroxidase (HRP) and chemically cross-linked to form Au NCs@BSA/HRP nanogels (ABH nanogels) with enhanced structural stability. The internal honeycomb architecture of the nanogels effectively prolongs the CL reaction. Within the nanogel, the dual catalytic system, composed of the Au NCs' mimetic peroxidase activity and the enzymatic activity of HRP, demonstrated high catalytic efficiency and stability. The nanogels also enhanced the luminol-H2O2 system, producing sustained glow-type luminescence. Leveraging these properties, a sensitive and convenient CL immunoassay for CA15–3 detection was developed, with ABH nanogels serving as the signal detector. The sensor exhibited a broad linear detection range from 0.005 to 100 U/mL and a detection limit of 0.0015 U/mL (S/N = 3).

A novel automated chemiluminescent enzyme immunoassay (CLEIA) for ADAMTS13 activity enables accompanying measurements of the inhibitory autoantibodies

A novel automated chemiluminescent enzyme immunoassay (CLEIA) for ADAMTS13 activity enables accompanying measurements of the inhibitory autoantibodies

Development of an automatic tubular chemiluminescence immunoassay using magnetic particles for the detection of antibodies against foot-and-mouth disease virus serotype A.

Foot-and-mouth disease virus (FMDV) causes significant financial losses in animal farms worldwide. Specific antibody monitoring of FMDV post-vaccination is important for evaluating vaccine efficacy, as well as the prevention and control of foot-and-mouth disease (FMD). In this study, we developed a fully automatic tubular chemiluminescence immunoassay method based on magnetic particles (A-MPCLIA) to specifically detect antibodies against FMDV serotype A. The diagnostic sensitivity (Dsn) and diagnostic specificity (Dsp) of A-MPCLIA were determined to be 96.05% and 99%, respectively, with a cut-off value of 30 activity units (U) by detecting the known background swine, cattle, and sheep serum. The developed method presented good Dsn, Dsp, and repeatability (CV < 10%). The activity units of A-MPCLIA, determined using 23 animals vaccinated with FMDV serotypes O, A, and Asia 1 univalent inactivated vaccine and nine serum samples from naïve animals, were significantly positively correlated with the titers of liquid-phase-blocking enzyme-linked immunosorbent assay (R2 = 0.992) and the titer of the virus neutralization test (R2 = 0.9577). In addition, A-MPCLIA requires less than 20 min, and it is fully automatic in detection. These results showed that A-MPCLIA is a suitable method for the detection of antibodies against FMDV serotype A in the future.

Prevalence of syphilis infection among migrant workers in Qatar: a nationwide cross-sectional survey

ObjectivesScant data are available on syphilis infection within migrant populations worldwide and in the population of the Middle East and North Africa region. This study investigated the prevalence of both...

SERS-based CRISPR/Cas12a assays for protein biomarker prostate-specific antigen detection.

Sensitive and accurate detection of protein biomarkers is crucial for disease diagnosis, especially for early diagnosis. Here, we describe surface-enhanced Raman scattering (SERS)-based CRISPR/Cas12a assays (S-CRISPR) for protein biomarker detection. Firstly, an S-CRISPR-driven enzyme-linked immunosorbent assay (S-CasLISA) was developed utilizing a capture antibody coated on a microplate to recognize the target and a detection antibody labeled with active DNA to trigger the activity of CRISPR/Cas12a. With this assay, we achieved detection of prostate-specific antigen (PSA) as models at thepicogram level. The limit of detection (LoD) of S-CasLISA was 0.17pgmL-1 and in the range of 0.1pgmL-1 to 10ngmL-1. Further, we applied aptamer to S-CRISPR (S-Apt-CRISPR), combining the high sensitivity of SERS with the high selectivity of aptamers, while simplifying the operation process of CRISPR detection of protein biomarkers. The proposed S-Apt-CRISPR also could detect picogram-level PSA and without repeated washing steps. The LoD of S-Apt-CRISPR was 0.35pgmL-1 and in the range of 0.1pgmL-1 to 10ngmL-1.Both SERS-based CRISPR/Cas12a assays were validated with clinical samples and demonstrated accuracy consistent with the chemiluminescence immunoassay. The introduction of the CRISPR/Cas12a system with SERS has the effect of improving the analytical capabilities of the system, thereby broadening and facilitating its application in the analysis of sensitive and accurate protein biomarkers.

Dynamics of the main parameters of carbohydrate metabolism and possible causes of its disorders in the autumn and winter periods in modern residents of the North

To assess the reserve capacity of the body from a physiological point of view, it is important to study the specific “northern” hormone metabolic profile of the body during critical (autumn and winter) periods in apparently healthy individuals born and living in the North.The aim of the work. To study the main parameters of carbohydrate metabolism, as well as possible causes of its disorders in the dynamics of autumn and winter periods in men living in the North.Materials and methods. The autumn (October) and winter (December) stages of the study included 45 men (mean age 40.0 ± 0.8 years) permanently residing in the Magadan Region. We used chemiluminescent immunoassay, enzymatic method and immunochromatographic assay.Results of the study. It was found that the average values of carbohydrate metabolism parameters in the examined male northern residents are comparable with the standard ranges with a shift towards higher values relative to the established limits and do not meet the main criteria of the “polar metabolic type”. “Polar metabolic type” is characterized by hypoglycemia and hypoinsulinemia against the background of elevated serum cortisol values. It is shown that during the critical period of the year from October to December (temperature transition through zero) against the background of relative “hypercortisolism”, activation of the insular apparatus of the pancreas is observed, accompanied by an increase in the insulin level, as well as the development of insulin resistance in the absence of compensatory secretion of β-cells of the pancreas. At the same time, the presence of signs of insulin resistance in northern male residents in the winter period may be determined by an imbalance towards greater dominance of the sympathetic nervous system, formed as a response to the critical period of the year.Conclusion. The obtained results indicate the formation of a transformed “northern” hormone metabolic profile of the body of a modern resident of the North, which should be considered as a certain adaptive response to the modern modification of the socio-economic lifestyle of northern residents (hypodynamia, overeating, etc.).

Correlation of Cytokine Profiles with Prostate-Specific Antigen and Disease Grade in Prostate Cancer.

BACKGROUND The development and progression of prostate cancer are multistep processes involving several growth factors, hormones, and cytokines. This study aimed to measure the serum concentrations of different cytokines and determine their correlation with prostate-specific antigen (PSA) levels and disease grade in patients with prostate adenocarcinoma. MATERIAL AND METHODS This cross-sectional study was conducted from March 2023 to March 2024 at the Clinic of Oncology of the University Hospital Center in Mostar, Bosnia and Herzegovina. Altogether, 50 male patients with prostate adenocarcinoma were included, of whom 28 had no proven metastases (PC group) and 22 had metastatic disease (MPC group). Serum concentrations of total (tPSA), free (fPSA), and complexed (cPSA) PSA were determined using a chemiluminescent microparticle immunoassay, whereas serum concentrations of cytokines were measured using a flow cytometry bead-based assay. RESULTS The MPC group had higher serum tPSA, fPSA, and cPSA levels than the PC group. The PC group had significantly higher serum levels of monocyte chemotactic protein (MCP)-1 than the MPC group (P=0.008). In the PC group, serum levels of interleukin (IL)-10 significantly correlated with cPSA. In the MPC group, serum concentrations of IL-1ß, tumor necrosis factor (TNF)-alpha, and IL-23 significantly correlated with disease grade. CONCLUSIONS Our study emphasizes the importance of MCP-1 in the development of prostate cancer, while IL-10 was the only cytokine whose serum level significantly correlated with cPSA. Serum concentrations of IL-1ß, TNF-alpha, and IL-23 may serve as potential biomarkers for disease grade.

Effect of UGT1A6 and UGT2B7 polymorphisms on the valproic acid serum concentration and drug-induced liver injury.

Aim: Valproic acid (VPA) is a classic broad-spectrum antiepileptic drug, with significant pharmacokinetic variability. Genetic polymorphisms contribute to this variability, influencing both VPA trough serum concentration (VPA concentration) and VPA-induced liver injury. Our study aims to investigate the association between polymorphisms of uridine diphosphate glucuronyl transferase (UGT) 1A6, UGT2B7 and VPA concentration and screen for potential genetic loci affecting VPA-induced liver injury.Methods: This study included epilepsy patients treated with VPA. PCR-RFLP method was used to determine the genotypes of UGT1A6 and UGT2B7. Chemiluminescent microparticle immunoassay was used to measure VPA concentration. Multiple linear regression and logistic regression were employed to analyze factors influencing VPA concentration and VPA-induced liver injury, respectively.Results: The correlation between UGT polymorphism and VPA concentration was analyzed in 133 samples. For VPA-induced liver injury, 105 patients were analyzed, with 29 in the liver injury group and 76 in the control group. Our finding showed patients with the UGT1A6-T19G variant had significantly lower VPA concentrations compared with wild-type patients and UGT1A6-T19G, A541G, A552C and UGT2B7-C802T, G211T, A268G polymorphisms showed no impact on VPA-induced liver injury.Conclusion: This study demonstrated UGT1A6-T19G polymorphisms affected the VPA concentration, providing a theoretical basis for the individualized clinical use of VPA.

Vitamin B12 concentrations vary greatly in milk donated to a large provincial milk bank, and are influenced by supplementation and parity

Feeding parent's milk with supplemental donor milk (DM) is the optimal way to feed very low birth weight (VLBW) infants instead of formula; however, suboptimal neurodevelopment persists. This is believed due, in part, to suboptimal nutrition. Given vitamin B12's role in neurodevelopment and increased adoption of plant-based diets among females of child-bearing age, we aimed to determine the adequacy of vitamin B12 in DM (n=380 donors) and associated donor characteristics. Vitamin B12 was measured in consecutive raw DM donations received at the milk bank from March 2020-2021 using a validated competitive chemiluminescent enzyme immunoassay (IMMULITE 2000, Siemens). Donor characteristics were obtained from screening records and associations with milk vitamin B12 concentrations explored using a generalized additive model. Donors were 32±4 years old (mean±SD), and DM was expressed 98±85 days postpartum. Vitamin B12 concentrations in DM had a median (25th, 75th percentile) of 232 (149, 373) pmol/L; 64% had concentrations <310pmol/L (common cut-off for inadequacy in healthy term-born infants). In a multivariable analysis, donors consuming a vitamin B12-containing supplement had higher DM vitamin B12 (β±SE: 80.3±25.4pmol/L; p=0.020) compared to those not taking a supplement. Primiparous donors had higher DM vitamin B12 than multiparous donors (36.7±18.2pmol/L greater; p=0.044). No associations were observed for other donor characteristics. Milk donated to a large human milk bank showed evidence of suboptimal vitamin B12; levels were associated with both donor vitamin B12-containing supplement use and parity. Further research as to whether and when milk bank donors are recommended to consume a supplement and the benefits and risks of routine vitamin B12 supplementation of DM-fed infants is warranted.

Effect of Iron Deficiency Anemia on Insulin Like- Growth Factor-1 Level Among Children: A Cross-Sectional Study

Objective: To assess the effect of iron deficiency anemia on the serum level of IGF-1 by comparison between low ferritin and low hemoglobin group and healthy control. Study Design: Comparative case control study. Place and Duration: Study The study was conducted at department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi, from Jul to Dec 2022. Methodology: The case control study was done in 110 children, aged between 2-14 years. Children with short stature and iron deficiency anemia (cases) were compared to a group of age and gender matched healthy children (controls). The sample size of 110 children was calculated using WHO sample size calculator. After brief history and examination, 3ml venous blood was drawn by standard technique. Complete blood picture estimation and the estimation of IGF-1 and ferritin levels was done. Parameters of complete blood picture included were Hemoglobin in (Hb), Red blood cells (RBC), Packed cell volume (PCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular volume (MCV). Data was analyzed using SPSS. Results: Comparison of the serum levels of IGF-1, analyzed by chemiluminescent immunoassay between two groups showed lower IGF-1 levels among iron deficiency anemia group (8.60 µmol/L) as compared to control group (38.80 µmol/L) which is statistically significant (p-value&lt;0.001). Results showed a positive correlation of Hemoglobin and serum ferritin with Serum IGF-1 levels with a r value of 0.78 and 0.81 (p-value 0.001) respectively. Conclusion: There is a strong correlation between Iron Deficiency anemia and IGF-1 levels. Children with Iron deficiency anemia have

Impact of gene polymorphisms involved in the vitamin D metabolic pathway on the susceptibility to and severity of autism spectrum disorder

This study explores the association between genetic variations in the vitamin D pathway and autism spectrum disorder (ASD) susceptibility and severity in Thai children. A total of 276 participants, including 169 children with ASD and 107 healthy controls, were recruited. Genotyping of vitamin D pathway genes (CYP2R1, CYP27B1, GC, and VDR) was conducted using TaqMan-based real-time PCR, while serum vitamin D levels were measured by chemiluminescence immunoassay. ASD severity was assessed via the Childhood Autism Rating Scale, 2nd Edition. Results reveal that the VDR gene (ApaI) rs7975232 is linked to a reduced ASD risk. In contrast, the GC gene rs7041 (A > C) polymorphism shows a significant association with increased ASD risk and severity, particularly in individuals with both the GC gene polymorphism and vitamin D insufficiency. Additionally, there was a higher prevalence of the GC1s isoform and GC1s-GC1s haplotype in children with ASD, associated with ASD severity. This study identified that individuals possessing GC rs7041 C alleles and the GC1s genotype (rs7041C/rs4588G) exhibit an increased susceptibility to and more severity of ASD. Further studies with larger cohorts are essential to fully understand these genetic polymorphisms’ roles.

Logistic regression analysis of risk factors for anxiety and depression in patients with coronary heart disease and subclinical hypothyroidism

We sought to explore the risk factors for anxiety and depression in patients with coronary heart disease and subclinical hypothyroidism through logistic regression analysis. A retrospective analysis was conducted on 168 patients with coronary heart disease and subclinical hypothyroidism admitted to the Department of Cardiology of our hospital from February 2020 to November 2022. Patients were categorized into the control group, anxiety group, and depression group based on the Hamilton Anxiety Scale (HAMA) and Hamilton Depression Rating Scale (HAMD) scores. All participants were informed about the protocol and provided signed informed consent upon inclusion. The study examined influencing factors for anxiety and depression in patients with coronary heart disease and subclinical hypothyroidism. Collect patients’ gender, age, presence or absence of chronic diseases (including Diabetes, hypertension and hyperthyroidism), sleep quality, dietary habits, psychosocial stress, living environment, social support, education level, and blood TSH levels. The linear relationship between anxiety, depression, and each influencing factor was quantified using the Pearson correlation coefficient. Blood level of TSH and free T4 were detected by chemiluminescence immunoassay. Multiple logistic regression was applied to analyze the factors influencing anxiety and depression in these patients. Various factors were identified as significant influencers of anxiety and depression in patients with coronary heart disease and subclinical hypothyroidism. For anxiety, presence or absence of chronic diseases, sleep quality, dietary habits, psychosocial pressure, living environment, and blood TSH levels were found to be influential (P < 0.05). Similarly, for depression, presence or absence of chronic diseases, sleep quality, social support, quality of life, social support, education level, and blood TSH levels were identified as significant factors (P < 0.05). The study revealed positive correlations between presence or absence of chronic diseases, psychosocial stress, and TSH levels with anxiety symptoms in patients with coronary heart disease and subclinical hypothyroidism (P < 0.05). Conversely, sleep quality, dietary patterns, and living environment showed negative correlations with anxiety symptoms (P < 0.05).Gender and age had no correlation with anxiety levels (P > 0.05). Presence or absence of chronic diseases and TSH levels were positively correlated with depressive symptoms in patients with coronary heart diseaseand subclinical hypothyroidism (P < 0.05). On the other hand, sleep quality, social support, quality of life, and educational level were negatively correlated with anxiety symptoms (P < 0.05). Gender and age had no correlation with depression (P > 0.05). Notably, TSH levels in both the anxiety and depression groups were higher than those in the control group (P < 0.05), with no significant difference in free T4 levels among the groups (P > 0.05). The combination of chronic illness types, living habits (sleep quality, dietary habits), psychosocial pressure, living environment, and TSH levels emerged as risk factors for anxiety in patients with coronary heart disease and subclinical hypothyroidism (P < 0.05). Similarly, the combination of chronic illness types, sleep quality, social support, quality of life, education level, and TSH levels were identified as risk factors for depression in these patients (P < 0.05). This logistic regression analysis underscores the significant impact of factors such as types of chronic illness, sleep quality, social support, living environment, education level, and TSH levels on anxiety and depression symptoms in patients with coronary heart disease and subclinical hypothyroidism. These findings highlight the importance of considering these multiple risk factors collectively when devising treatment and management strategies to reduce the risk of mental health issues in this patient population.

Abstract A059: Serum EphA2 proteolytic fragment is a potent biomarker for diagnosing a very early stage of ductal pancreatic carcinoma

Abstract Erythropoietin-producing hepatocellular receptor A2 (EphA2) is a receptor tyrosine kinase, and its expression increases with malignant progression of cancer. Therefore, EphA2 is believed to be a candidate for molecular cancer therapy and many candidate drugs targeting EphA2 have been developed, but their effects are limited. The reason is its complex signaling functions. In fact, EphA2 has a tumor suppressor function regulated by the stimulation of the Ephrin-A ligand, while in the absence of ligand stimulation, it has a prooncogenic function in collaboration with EGFR and its downstream signaling molecules. The opposite functions of EphA2 make it difficult to develop it as a cancer drug target. We have previously demonstrated that a type-1 matrix membrane metalloprotease (MT1-MMP) at cancer cell surface interacted with EphA2 and selectively cleaved its N-terminal ligand binding site, converting it into a prooncogenic signaling transmitter (Koshikawa N et al., Cancer Research, 2015). We also found that the N- terminal fragment of EphA2 (termed EphA2-NF) released by MT1-MMP cleavage increased in sera from patients with gastric cancer and ductal pancreatic carcinoma (PDC) (Koshikawa N et al., Cell Death &amp; Disease, 2017). In this study, we compare the expression of EphA2-NF in serum from a healthy donor (HD: 150 cases), patients with chronic pancreatic disease (87 cases), and PDC (578 cases) by automated chemiluminescence immunoassay with a specific EphA2-NF monoclonal antibody. As a result, PDC cases, including stage I/II PDC, show significantly higher levels of EphA2-NF than HD (Sato et al. Cancer Research Communications, 2023). Furthermore, PDC occurred in the intraductal papillary mucinous neoplasm (IPMN) cases with high levels of EphA2-NF during follow-up observation. Therefore, to verify that the EphA2 proteolytic cleavage occurred in the PDC onset, we analyzed 50 surgical resected tissues from patients with PDC and IPMN to detect the N and C-terminal regions of EphA2 by immunohistochemical staining. Interestingly, PDC, including early stage PDC cases, lacked the N-terminus of EphA2. Furthermore, the deletion of N-terminus of EphA2 was observed in all tested cases pathologically diagnosed as intraductal papillary mucinous adenocarcinoma (IPMC) (Sato S et al., Cancer Research Communications, 2023). These results suggest that serum EphA2-NF has the potential to revolutionize the early detection and management of PDC, providing a valuable tool to diagnose the disease in very early stages when treatment results may be more favorable. Collaborators: Masatoshi Nakagawa, Eisaku Yoshida and Toru Yoshimura (Research and Development, Abbott Japan LLC) Citation Format: Naohiko Koshikawa, Shinya Sato, Kouki Nio, Takeshi Terashima, Makoto Ueno, Taro Yamashita. Serum EphA2 proteolytic fragment is a potent biomarker for diagnosing a very early stage of ductal pancreatic carcinoma [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A059.

Abstract 4134234: Association Between the Natriuretic Peptides-Cyclic Guanosine Monophosphate Cascade and Left Ventricular Reverse Remodeling After Acute Anterior Myocardial Infarction

Background: Natriuretic peptides and their second messenger, cyclic guanosine monophosphate (cGMP), play a critical role in the pathogenesis of cardiovascular diseases by inhibiting the renin-aldosterone system, cardiac hypertrophy, and fibrosis. Recently developed chemiluminescent enzyme immunoassays for proBNP have enabled the estimation of mature BNP (emBNP) by calculating the difference between total BNP and proBNP. Research Questions: The role of cGMP in left ventricular remodeling after acute myocardial infarction (AMI) remains poorly understood. Aims: We aimed to investigate the relationship between the natriuretic peptide-cGMP cascade and left ventricular reverse remodeling (RR) in patients with anterior AMI. Methods: In a prospective 67 patients experiencing their first anterior AMI, plasma BNP fractions and cGMP levels were measured at immediate primary percutaneous coronary intervention (PPCI), 3 days, and 3 months post-AMI. Change in left ventricular end-systolic index (delta-LVESVI) was calculated as the difference between median 10 months and day 10 using cardiac magnetic resonance imaging. Patients were divided into RR and non-RR groups based on regression analysis to compute residuals representing the differences between each individual’s observed delta-LVESVI and delta-LVESVI adjusted by plasma pro-BNP concentration at immediate PPCI because of an inverse correlation between delta-LVESVI and plasma pro-BNP concentration at immediate PPCI (r=0.243, p=0.047). RR group (n=33) demonstrated below-median residual delta-LVESVI, while non-RR group (n=34) had above-median residual. Results: RR group exhibited significantly higher freedom from major adverse cerebral and cardiovascular events (MACCE) during a median of 8.1-year (Interquartile range: 6.7–8.8) follow-up (P=0.024) (Figure A) . Multivariable analysis identified logarithmic plasma pro-BNP, and emBNP at day 3 post-AMI, and plasma cGMP levels at immediate PPCI and day 3 post-AMI, as independent predictors of RR (P&lt;0.05) even after adjusting age, male, hypertension, and infarct size. (Figure B-D) . Conclusion: Early-phase activation of the natriuretic peptides-cGMP cascade may play a protective role in left ventricular remodeling in patients with anterior AMI.

D-hypovitaminosis in menopausal disorders clinic: Correction possibilities

Introduction. The postmenopausal period being a significant part of a modern woman’s life. The persistence of menopausal disorders clinic can be combined with the hypovitaminosis D state, which, in turn, can further aggravate the existing symptoms. Vitamin D supplementation may be one of the pharmacological approaches to correct menopausal disorders.Aim. To evaluate the contribution of hypovitaminosis D to the clinic of menopausal disorders; to determine the clinical efficacy of using cholecalciferol in late postmenopausal residents of Yekaterinburg.Materials and methods. During the period from 2018 to 2021, 144 late postmenopausal patients from 56 to 79 years old living in Yekaterinburg and do not need outside help in everyday life took part. The following parameters of anthropometry, the severity of modified menopausal index (MMI), hospital anxiety and depression scales (HADS), Mini-Mental State Examination (MMSE), an assessment of serum 25(OH)D level (chemiluminescent immunoassay; Access 2, Beckman Coulter, USA) were studied. Clinical efficacy of cholecalciferol was evaluated in 68 patients in the prospective cohort study design. Micellated water-soluble form of cholecalciferol in standard doses recommended by the Russian Association of Endocrinologists (RAE) was used.Results. Vitamin D deficiency and insufficiency are associated with increased parameters of body weight (p = 0.048), waist circumference (p = 0.018) and hips (p = 0.016), increased cases of decreased performance (p = 0.046) and subclinical symptoms anxiety (p = 0.033). Supplementation of standard therapeutic and maintenance doses of cholecalciferol for 6 months contributed to a significant reduction in neurovegetative component of modified menopausal index in 54.4% of cases (p = 0.035) and improvement in cognitive function parameters in 50.0% of cases (p = 0.023) in late postmenopausal patients.Conclusions. The use of cholecalciferol in adequate doses can be considered as an important component of complex therapy for women in late postmenopausal period.

Fabrication of functional interface on magnetic beads via various amino acids and their application in chemiluminescent immunoassay as carrier

Magnetic polymer microspheres with superparamagnetism, high specificity, and monodispersity play a crucial role in the field of in vitro diagnostics. However, the surface modification process of magnetic beads is often complex, and it remains a significant challenge to prepare high-performance magnetic beads easily. To overcome these drawbacks, herein we fabricated functional interface on magnetic bead with the various amino acid via the ring-opening reaction of amino acids with epoxy groups, with attempt to produce carboxylated magnetic beads (MPS-GA) in a convenient way. Results indicate that when compared to other amino acids, the phenylalanine magnetic beads (MPS-GA1) developed in this study exhibit strong adsorption for mouse immunoglobulin (IgG), streptavidin (SA), and protamine (PA), with an IgG adsorption capacity of 53.5 μg/mg and a coupling capacity of 52.5 μg/mg. It is found that electrostatic forces and hydrophobic interactions are key factors influencing biomolecular interactions. Additionally, these magnetic beads can generate strong chemiluminescent signals, significantly reducing background levels by up to 99.7 %. Therefore, the magnetic beads proposed in this paper can serve as carriers for chemiluminescent immunoassay (CLIA), providing new insights into the synthesis of high-quality magnetic bead.

Invisible burden: Prevalence of Chagas disease in Latin American migrants in Turin, North-Western Italy – A STROBE compliant-cross-sectional study

ObjectivesChagas disease (CD), or American trypanosomiasis, is a parasitic disease caused by Trypanosoma cruzi, primarily transmitted by triatomine bugs. Increased travels and migrations introduced CD to non-endemic regions, including Europe. In Italy, the disease has raised public attention mainly in northern regions, where Latin American migrant population is larger.This study aims to describe a CD screening initiative in Turin, Piedmont, during the World Chagas disease Day. We focused on migrants from CD endemic countries of Latin America. MethodsA total of 63 serum samples from Latin American migrants were collected and tested for CD with a chemiluminescence immunoassay (CLIA) for initial screening, followed by Western Blot (WB) for confirmation, analyzing demographic data. ResultsOut of the 63 individuals screened, two tested positive for CD. Both positive cases were from Bolivia. Seropositive individuals were 54 and 72 years-old, and lived in Bolivia for 31 and 69 years. ConclusionsThe screening initiative suggests an underestimated prevalence of CD among Latin American migrants in Turin, underscoring the need of targeted public health interventions and emphasizing the importance of CD screening and awareness programs.